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PXD051862

PXD051862 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleA simple and efficient approach for preparing cationic coating with tunable electroosmotic flow for capillary zone electrophoresis-mass spectrometry-based top-down proteomics
DescriptionCapillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has become a valuable analytical technique in top-down proteomics (TDP) in recent years. CZE-MS/MS-based TDP typically employs separation capillaries with neutral coatings (i.e., linear polyacrylamide, LPA). However, issues related to separation resolution and reproducibility remain with the LPA-coated capillaries due to the unavoidable non-specific protein adsorption onto the capillary wall. Cationic coatings can be critical alternatives to LPA coating for CZE-MS/MS-based TDP due to the electrostatic repulsion between the positively charged capillary inner wall and proteoform molecules in the acidic separation buffer. Unfortunately, there are only very few studies using cationic coating-based CZE-MS/MS for TDP studies. In this work, we aimed to develop a simple and efficient approach for preparing separation capillaries with cationic coating, i.e., poly(acrylamide-co-(3-acrylamidopropyl)trimethylammonium chloride, PAMAPTAC for CZE-MS/MS-based TDP. The PAMAPTAC coating-based CZE-MS produced significantly better separation resolution of proteoforms compared to traditionally used LPA-coated approach. It achieved reproducible separation and measurement of simple proteoform mixture and a complex proteome sample (i.e., a yeast cell lysate) regarding migration time, proteoform intensity, and the number of proteoform identifications. The PAMAPTAC coating-based CZE-MS enabled the detection of large proteoforms (≥30 kDa) from the yeast cell lysate reproducibly without any size-based prefractionation. Interestingly, the electrophoretic mobility of proteoforms using the PAMAPTAC coating can be predicted accurately using a simple semiempirical model. The results render the PAMAPTAC coating as a valuable alternative to the LPA coating to advance CZE-MS-based TDP towards high-resolution separation and highly reproducible measurement of proteoforms in complex samples.
HostingRepositoryPRIDE
AnnounceDate2024-10-15
AnnouncementXMLSubmission_2024-10-15_05:21:36.570.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterLiangliang Sun
SpeciesList scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationListphosphorylated residue; acetylated residue; monohydroxylated residue
InstrumentQ Exactive HF; Orbitrap Exploris 480
Dataset History
RevisionDatetimeStatusChangeLog Entry
02024-04-29 14:36:15ID requested
12024-10-15 05:21:37announced
Publication List
Wang Q, Gao G, Fang F, Wang Q, Lundquist PK, Sun L, A simple and efficient approach for preparing cationic coating with tunable electroosmotic flow for capillary zone electrophoresis-mass spectrometry-based top-down proteomics. Anal Chim Acta, 1328():343162(2024) [pubmed]
10.1016/j.aca.2024.343162;
Keyword List
submitter keyword: Top-down proteomics
Proteoform
Capillary zone electrophoresis-mass spectrometry
Covalent cationic coating
Reproducibility
large proteoforms
Contact List
Liangliang Sun
contact affiliationDepartment of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI 48824, USA
contact emaillsun@chemistry.msu.edu
lab head
Liangliang Sun
contact affiliationMichigan State University
contact emaillsun@chemistry.msu.edu
dataset submitter
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Dataset FTP location
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